Background
End-stage renal disease (ESRD) patients usually develop extensive and progressive vascular calcification, and lots of calcification inhibitors as well as pro-calcifying factors are involved in the process. However, the mechanisms of vascular calcification in ESRD patients are still ill-defined.
Methods
The participants in the study were selected at the Second Xiangya Hospital, Central South University, and the plasma exosomes derived from ESRD patients (ESRD-Ex), renal transplant recipients (RTR-Ex), and the normal health control group (Nor-Ex) were isolated by ExoQuick-TC kit. Transmission electron microscopy and molecular size analysis were used to assess the morphology and size of exosomes. Alizarin Red S staining was carried out to detect calcification of vascular smooth muscle cells (VSMCs). Protein levels of Fetuin-A, matrix gla protein (MGP), Annexin-A2, bone morphogenetic protein (BMP-2), and receptor activator for nuclear factor-κ B ligand (Rankl) were measured by Western Blot analysis and the contents of that were detected using ELISA. Coronary artery calcification scores (CACS) were quantified via Agaston and analysed by Siemens CaScoring software.
Results
Compared with Nor-Ex, the ESRD-Ex promoted calcification of VSMCs significantly, and RTR-Ex could attenuate VSMCs calcification. Moreover, the protein concentration of ESRD-Ex was significantly higher than Nor-Ex and RTR-Ex, and the content of both MGP and Fetuin-A, the calcification inhibitors, were prominently lower in ESRD-Ex than those in Nor-Ex. The content of Annexin-A2, one of the calcification promoters, was significantly higher in ESRD-Ex and RTR-Ex than that in Nor-Ex. But, BMP-2 and Rankl had no significant difference among the three groups. In addition, the content of Fetuin-A in RTR-Ex was higher than that in ESRD-Ex, though it was still lower than in Nor-Ex. Furthermore, the content of both plasma Fetuin-A and MGP were negatively while that of Annexin-A2 was negatively correlated to CACS.
Conclusions
These results indicated that ESRD-Ex significantly promoted VSMCs calcification while renal transplantation could partially attenuate the effect of exosomes. Fetuin-A and MGP were decreased but Annexin-A2 was increased in ESRD-Ex, and renal transplantation could increase the level of Fetuin-A rather than MGP.